PMID- 8144933
OWN - NLM
STAT- MEDLINE
DCOM- 19940429
LR  - 20071114
IS  - 0022-1767 (Print)
IS  - 0022-1767 (Linking)
VI  - 152
IP  - 7
DP  - 1994 Apr 1
TI  - Biochemical and biologic characterization of murine monocyte chemoattractant 
      protein-1. Identification of two functional domains.
PG  - 3541-9
AB  - Monocyte chemoattractant protein-1 (MCP-1) is a member of the chemokine-beta (or 
      C-C) family of cytokines. Murine MCP-1, first identified as the JE gene, differs 
      from human MCP-1 in molecular size and extent of glycosylation. We have used 
      Chinese hamster ovary cells to express recombinant murine MCP-1 and find that its 
      predominant form is a microheterogeneous protein of M(r) approximately 25,000. 
      Most of MCP-1's microheterogeneity is due to variable amounts of sialic acid that 
      are terminally attached to a constant number of O-linked oligosaccharide chains 
      per molecule. This carbohydrate, along with a small amount of N-linked 
      carbohydrate, accounts for 50% of the apparent molecular size of murine MCP-1 and 
      is not required for in vitro monocyte chemoattractant activity. Mutational 
      analysis shows that most of the carbohydrate is added to a 49-amino acid 
      C-terminal domain that is not present in human MCP-1 and is not required for in 
      vitro biologic activity, suggesting that murine MCP-1 consists of an N-terminal 
      domain containing monocyte chemoattractant activity and a heavily glycosylated 
      C-terminal domain of as yet unknown function. MCP-1 produced in COS cells 
      contains a small amount of sulfate, but Chinese hamster ovary-produced MCP-1 does 
      not. The absence of sulfate does not alter MCP-1's in vitro chemoattractant 
      properties. In vitro, highly purified murine MCP-1 attracts monocytes, but not 
      neutrophils, with a specific activity similar to human MCP-1 (EC50 approximately 
      0.5 nM). Equilibrium binding experiments with human monocytes reveal the presence 
      of approximately 3000 binding sites per cell with a Kd of 0.77 nM. In vivo, 
      injection of up to 1 micrograms murine MCP-1 in a variety of murine strains 
      induces the appearance of a sparse mixed inflammatory infiltrate. The disparity 
      between MCP-1's in vitro and in vivo effects suggests that other factors may be 
      required to elicit a full-blown monocyte chemotactic response to MCP-1 in vivo.
FAU - Ernst, C A
AU  - Ernst CA
AD  - Department of Medicine, Dana-Farber Cancer Institute, Harvard Medical School, 
      Boston, MA 02115.
FAU - Zhang, Y J
AU  - Zhang YJ
FAU - Hancock, P R
AU  - Hancock PR
FAU - Rutledge, B J
AU  - Rutledge BJ
FAU - Corless, C L
AU  - Corless CL
FAU - Rollins, B J
AU  - Rollins BJ
LA  - eng
GR  - CA53091/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Immunol
JT  - Journal of immunology (Baltimore, Md. : 1950)
JID - 2985117R
RN  - 0 (Chemokine CCL2)
RN  - 0 (Chemotactic Factors)
RN  - 0 (DNA Primers)
RN  - 0 (Glycoproteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Sulfates)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Chemokine CCL2
MH  - Chemotactic Factors/*chemistry
MH  - Chemotaxis, Leukocyte
MH  - DNA Primers/chemistry
MH  - Glycoproteins/chemistry
MH  - Glycosylation
MH  - Humans
MH  - Mice
MH  - Molecular Sequence Data
MH  - Monocytes/metabolism
MH  - Recombinant Proteins
MH  - Sulfates/chemistry
EDAT- 1994/04/01 00:00
MHDA- 1994/04/01 00:01
CRDT- 1994/04/01 00:00
PHST- 1994/04/01 00:00 [pubmed]
PHST- 1994/04/01 00:01 [medline]
PHST- 1994/04/01 00:00 [entrez]
PST - ppublish
SO  - J Immunol. 1994 Apr 1;152(7):3541-9.